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首页> 外文期刊>Plant Cell Reports >Isolation and characterization of a molecule stimulatory to growth of somatic embryos from early stage female gametophyte tissue of loblolly pine
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Isolation and characterization of a molecule stimulatory to growth of somatic embryos from early stage female gametophyte tissue of loblolly pine

机译：松树早期雌配子体组织中刺激体细胞胚生长的分子的分离和表征

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Loblolly pine (LP, Pinus taeda) is the primary commercial species in southern forests of the US. Somatic embryogenesis (SE) is an effective technique to implement clonal tree production of high-value genotypes from breeding and genetic engineering programs. Unlike angiosperm embryos with attached cotyledons as seed storage organs, the diploid conifer embryo is surrounded by the unattached haploid female gametophyte (FG). The FG is not present in culture. This presents a dilemma if the FG produces necessary or regulatory compounds for embryo growth, since in culture these important compounds would be missing and would have to be added as supplements. We report here the direct evidence that extracts from early-stage FG indeed stimulate early-stage somatic embryo (SME) growth and multiplication, whereas extracts from late-stage FG inhibit early-stage SME growth. Furthermore, we have now isolated this stimulatory substance from early-stage FG tissue, and identified this substance as citric acid on the basis of NMR and mass spectrometry. We then demonstrated that topical application of citric acid to SMEs stimulates embryo colony growth at P = 0.05. Moreover, we find that there is a good correlation between the amount of citric acid isolated from FG tissue (65 nmoles per stage 2-3 FG) and the amount of citric acid that stimulates colony growth (25-50 nmoles) when applied topically to SMEs. This approach of isolating and characterizing a molecule from plant tissue, and investigating its role on SE processes can provide valuable information leading to further applications of these molecules to improve LP SE protocols.
机译：火炬松（LP，Pinus taeda）是美国南部森林中的主要商业树种。体细胞胚发生（SE）是一种有效的技术，可通过育种和基因工程程序实现高价值基因型的无性系生产。与以子叶为种子贮藏器官的被子植物胚不同，二倍体针叶树胚被未附着的单倍体雌配子体（FG）包围。 FG在文化中不存在。如果FG产生胚胎生长所必需的或调节性的化合物，这将是一个难题，因为在培养中这些重要的化合物会丢失，必须作为补充添加。我们在这里报告的直接证据表明，早期FG的提取物确实刺激了早期体细胞胚（SME）的生长和繁殖，而晚期FG的提取物抑制了早期SME的生长。此外，我们现在已经从早期FG组织中分离出这种刺激性物质，并根据NMR和质谱法将该物质鉴定为柠檬酸。然后，我们证明了柠檬酸对中小企业的局部应用可刺激胚胎菌落的生长，P = 0.05。此外，我们发现从FG组织中分离出的柠檬酸量（每个阶段2-3 FG为65 nmoles）与局部应用柠檬酸刺激菌落生长的柠檬酸量（25-50 nmoles）之间存在良好的相关性。中小企业。这种从植物组织中分离和表征分子，并研究其在SE过程中的作用的方法可提供有价值的信息，从而可进一步应用这些分子来改善LP SE方案。

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